Human Activity-Optimized CRISPR Knockout Library (3 sub-libraries in lentiCRISPRv1)
(Pooled Library #1000000100)

Library Details

• Species: Human
• Genes targeted: 18,663
• gRNAs: 187,535
• Controls: 1000 non-targeting, 500 intergenic
• Lentiviral Generation: 3rd

Library Shipping

This library will be delivered as three pooled DNA sub-libraries in microcentrifuge tubes on blue ice. The tubes' contents will not necessarily be frozen. For best results, minimize freeze-thaw cycles.

Note that each subpool must be transformed separately.

You will receive two sub-libraries with 90,000 gRNAs each at

• Volume: ∼15 µL
• Concentration: 10 ng/µL

You will additionally receive one sub-library (hL3C9) with 5,000 gRNAs each at

• Volume: ∼20 µL
• Concentration: 20 ng/µL

Resource Information

• Protocols:
  • Library Transformation Protocol (15.1 KB)
  • Screening Protocol (15.8 KB)
  • PCR protocol for Illumina sequencing preparation (14.3 KB)
• Depositor Data:
  • List of targeted genes and sgRNA sequences (10.0 MB)
  • Sequencing data (sgRNA read counts for each subpool) (2.7 MB)
  • Data for hL3C9 (11 MB)
• Terms and Licenses:
  • UBMTA

Depositor Comments

The depositing lab notes that the DNA preps should be performed separately and then mixed together during virus production at ratios consistent with the pool size.

Diagram of Library Screening Process

When performing sgRNA validation in a Cas9-expressing cell line, the depositing laboratory recommends using plasmid pLenti-sgRNA (Addgene #71049).

sgRNA diversity after initial amplification

sgRNA diversity after infection of a target cell population